Search

The cDNA sequence of a small heat shock protein ( hsp19.7) was cloned and sequenced from the rice stem borer, Chilo suppressalis Walker. The cDNA encoded a protein of 177 amino acids with a calculated molecular weight of 19.7 kDa. The deduced amino acid sequence showed the highest identity of 90% to Bombyx mori hsp19.9. Expression levels of hsp19.7 were similar between diapausing and non-diapausing larvae. In non-diapausing larvae, but not in diapausing ones, hsp19.7 expression was upregulated by cold acclimation. Involvement of hsp19.7 in larval diapause and cold tolerance in C. suppressalis is discussed. [ABSTRACT FROM AUTHOR]

The present work reports the results of a study on the isolation and characterization of matrix polysaccharides in the cell walls of galls formed by an aphid ( Neothoracaphis yanonis) on Distylium racemosum leaves. Cell walls were isolated from both healthy Distylium leaf and gall tissues and then extracted sequentially with cyclohexane- trans-1,2-diaminetetra-acetate (CDTA), Na2CO3, 1 m KOH, and 4 m KOH. The amount of pectin solubilized from gall cell walls was approximately 2.6-fold higher than the pectin solubilized from leaf cell walls, whereas the amount of hemicellulose solubilized from gall cell walls was 1.4-fold higher than that from normal leaf cell walls. When the polysaccharides were fractionated by anion-exchange chromatography, considerable increases in arabinose and galactose were observed in CDTA-soluble pectic polymer (fraction PI-1) from gall cell walls, whereas the gall cell walls had less xylose in 1 m KOH-soluble hemicellulosic polymers (fractions HI-2, HI-3, and HI-4) than did the cell walls from the healthy leaf. The hemicellulosic polymers of the gall cell walls exhibited distinctly different patterns of molecular mass, compared with the healthy leaf cell walls. These results suggest that an extensive change occurs in the matrix polysaccharide structure of the cell walls of Distylium galls formed by an aphid. In addition, many glycosylhydrolase activities were detected in the protein fraction solubilized with strong saline solution from the gall cell walls, and the activities of β-galactosidase, β-xylosidase and α- l-arabinofuranosidase were considerably increased under gall formation. [ABSTRACT FROM AUTHOR]

Presents information on a study that investigated the effects of induced apoptosis by a polyamine synthesis inhibitor, methyl-glyoxal bis (cyclopentylamidinohydrazone) in cultured rheumatoid synoviocytes in vitro and rheumatoid synovial tissue. Methodology of the study; Results and discussion on the study.

A gene encoding heat shock transcription factor (HSF) was cloned and sequenced from cultured cells of the cabbage armyworm, Mamestra brassicae. The cDNA potentially encoded a 699-aa protein, with a calculated molecular weight of 77.8 kDa. Deduced amino acid identities to HSFs from Aedes aegypti and Drosophila melanogaster were 36 and 34%, respectively. Analysis of the genomic DNA revealed eight exons and three optional exons: a, b, and c. Exon a contained a premature in-frame stop codon that would generate a truncated protein. When the cells were exposed to high temperature or cadmium, no particular alternative transcripts showed significant up- or down-regulated expression relative to the total amount of the transcripts. These results suggest that alternative splicing may not be a principal mechanism for regulation of M. brassicae HSF gene expression in response to heat shock and cadmium., ((c) 2009 Wiley Periodicals, Inc.)

Cold hardiness of the freshwater apple snail, Pomacea canaliculata, varies seasonally. We investigated lethal factors and physiological changes arising from exposure of P. canaliculata to low temperatures. Snails did not survive freezing. The supercooling point of cold-acclimated (cold tolerant) snails (-6.6+/-0.8 degrees C) did not differ significantly from that of non-acclimated ones (-7.1+/-1.5 degrees C) under laboratory conditions. Furthermore, snails died even under more moderately low temperatures approaching 0 degrees C. These results indicate that indirect chilling injury is a factor in the death of P. canaliculata at low temperatures. Regardless of whether the snails were acclimated to low temperatures, all of the dead, and even some of the snails still alive at 0 degrees C, had injured mantles, indicating that the mantle may be the organ most susceptible to the effects of low temperatures. The concentration of glucose in the posterior chamber of the kidney and concentration of glycerol in the digestive gland were significantly higher in cold-acclimated snails than in non-acclimated ones, suggesting carbohydrate metabolic pathways are altered in snails during cold acclimation.

We examined the genomic organization of the para-sodium channel alpha-subunit gene of the diamondback moth, Plutella xylostella (L.). The nucleotide sequence contained 34 putative exons, which covered almost the entire coding region of the gene producing 1,889 amino acid residues. Deduced amino acid identity to the hscp locus of Heliothis virescens was 84%. Comparison of deduced amino acid sequences of the permethrin-resistant and -susceptible strains showed two substitutions other than kdr and super-kdr-like substitutions. They were Ala to Thr (A1060T) and Pro to Ser (P1836S) at the linker region of the domains II-III and the carboxyl terminus, respectively. Furthermore, we developed PCR amplification protocols for the rapid detection of both substitutions., (Copyright 2008 Wiley-Liss, Inc.)

The frequencies of the L1014F and T929I mutations, both of which are involved in nerve insensitive resistance to pyrethroids, were examined in field and laboratory strains of the diamondback moth, Plutella xylostella at DNA and RNA levels. Results showed that the resistance allele frequencies at the L1014F and T929I sites in the field strains were respectively, 82.8-100% and 72.9-94.4%. No posttranscriptional regulation of the L1014F mutation was observed. The examined insects were classifiable into four groups according to the expression patterns of mutually exclusive exons 18a and 18b. Most insects in the field strains expressed transcripts containing exon 18b more abundantly than those containing exon 18a, although both transcripts were expressed with similar proportions in all insects of the laboratory strains. Some other insects expressed a chimeric transcript comprising parts of exons 18a and 18b. Deduced amino acid sequences of the chimeric transcript encoded amino substitution from Met to Ile at the site corresponding to the super-kdr mutation (M918T) in Musca domestica. The frequencies of the M918I mutation in the field strains were 5.0-19.4%. Analyses of the genomic organization revealed that the chimeric sequences are encoded in the genome.

The apple snail, Pomacea canaliculata, is an invasive freshwater snail. It increases its cold hardiness before winter. However, the physiological mechanism of cold hardiness in molluscs is poorly understood, especially in freshwater molluscs. In this study, we examined the changes in low molecular weight compounds, glycogen and lipids, in the body of P. canaliculata in association with the development of cold hardiness. When snails without cold hardiness were experimentally cold-acclimated, the amount of glycerol, glutamine, and carnosine increased, while glycogen and phenylalanine decreased. Overwintering cold-tolerant snails collected from a drained paddy field in November also showed increased glycerol in their bodies with decreasing glycogen concentration, compared to summer snails collected from a submerged field. Water content also decreased during the cold acclimation, although the water loss was minimal. These results indicate that the freshwater snail, P. canaliculata enhances cold hardiness by accumulation of some kinds of low molecular weight compounds in its body as some insects do. However, the actual function of each low molecular compound is still unknown.

We cloned and characterized cDNA of two storage protein (SP) genes, PxSP1 and PxSP2, from the diamondback moth, Plutella xylostella (Lepidoptera: Yponomeutidae) and investigated their expression. PxSP1 and PxSP2 each encoded a putative protein of 91 kDa. Nucleotide and deduced amino acid identities between the two genes were 79% and 82%, respectively. Amino acid composition (methionine>4%), sequence homology with other insect storage proteins and the phylogenetic analysis suggested that the genes belong to the subfamily of moderately methionine-rich SP genes. The genes were predominantly expressed in the last instar female larvae and the mRNA levels were suppressed by treatment with a juvenile hormone-analog. Treatment of female larvae with sublethal dose of a pyrethroid caused a significant increase in mRNA levels of both genes. Induction of PxSP1 and PxSP2 genes as a result of pyrethroid application may have implications with respect to reproduction as methionine-rich proteins are known as a key element for egg production.

Heat shock protein (HSP) genes, hsp90, hsp70, hsc70, hsp20.7, and hsp19.7, were cloned and sequenced from cultured cells of the cabbage armyworm, Mamestra brassicae. Analyses of the cDNA sequences revealed open reading frames of 2,151, 1,914, 1,962, 540, and 465 bp in lengths, which encode proteins with calculated molecular weights of 82.5, 69.9, 71.6, 20.7, and 19.7 kDa, respectively. An increased expression was observed in all five genes after exposure to a high temperature. The induction of gene expression was not observed during a low temperature exposure, but was observed when the cells were recovered at ambient temperature. Expression of hsp90, hsp70, and hsp20.7 was induced after exposure to 2 microM of cadmium, while the minimum cadmium concentration for induction of hsp19.7 was 5 microM. The induction of hsp90 expression was somewhat masked by basal levels of expression. Only hsp20.7 expression was induced by exposure to copper. Lead did not induce expression of any of the HSP genes tested. Cadmium-induced up-regulation of hsp70 expression was lasted longer than heat-induced one. These results suggest that hsp70 could be useful to assess the cellular distress or injury induced by cadmium.

Overwintering freeze-tolerant larvae of Chilo suppressalis can survive at -25 degrees C, but non-diapausing larvae cannot. We reported earlier that to prevent intracellular freezing, which causes death in overwintering larvae of the Saigoku ecotype distributed in southwestern Japan, water leaves and glycerol enters fat body cells through water channels during freezing. However, it is still unclear how diapause and low-temperature exposure are related to the acquisition of freeze tolerance. We compared the extent of tissue damage, accumulation of glycerol, and transport of glycerol and water in fat body tissues between cold-acclimated and non-acclimated non-diapausing and diapausing larvae. The tissue from cold-acclimated diapausing larvae could survive only when frozen in Grace's insect medium with 0.25 M glycerol at -20 degrees C. The protection provided by glycerol was offset by mercuric chloride, which is a water-channel inhibitor. Fat body tissue isolated from non-acclimated diapausing larvae was injured by freezing even though glycerol was added to the medium, but the level of freezing injury was significantly lower than in non-diapausing larvae. Radiotracer assays in cold-acclimated diapausing larvae showed that during freezing, water left the cells into the medium and glycerol entered the cells from the medium at the same time. Therefore, in Saigoku ecotype larvae of the rice stem borer, both diapause and cold-acclimation are essential to accumulate glycerol and activate aquaporin for the avoidance of freezing injury.

Helicoverpa armigera (Hübner) exhibits a facultative pupal diapause, which depends on temperature and photoperiod. Pupal diapause is induced at 20 degrees C by short photoperiods and inhibited by long photoperiods during the larval stage. However, in some pupae (35% of males and 57% of females) of a non-selected field population from Okayama Prefecture (34.6 degrees N), diapause is not induced by short photoperiods. In the present experiment, the importance of temperature for diapause induction was studied in the non-diapausing strain, which was selected from such individuals reared at 20 degrees C under a short photoperiod of 10L:14D. Furthermore, the sensitive stage for thermal determination of pupal diapause was determined by transferring larvae of various instars and pupae between 20 degrees C and 15 degrees C. Diapause was induced by 15 degrees C without respect to photoperiod. When larvae or pupae reared from eggs at 20 degrees C under a short or a long photoperiod were transferred to 15 degrees C in the periods of the middle fifth instar to the first three days after pupation, the diapause induction rate was significantly reduced in both males and females, especially in females. In contrast, when larvae or pupae reared at 15 degrees C were transferred to 20 degrees C in the same periods, diapause was induced in males, but not in females. However, the diapause induction rate of pupae transferred to 20 degrees C on the fourth day after pupation was significantly increased in females. The results show that temperature is the major diapause cue in the photoperiod-insensitive strain and the periods of middle fifth larval instar to early pupal stage are the thermal sensitive stages for pupal diapause induction with some different responses to temperatures between males and females in H. armigera.

We cloned and characterized two hexameric storage protein genes, PxAry1 and PxAry2, from Plutella xylostella and investigated the expression pattern in different developmental stages and in response to treatment by a juvenile hormone (JH) analog. The complete coding sequences of PxAry1 and PxAry2 are comprised of 2,097 and 2,094 bp with 699 and 698 amino acid residues, respectively. Signal peptides of 16 amino acids are predicted at the N-termini. According to both the phylogenetic analysis and amino acid composition (>16% aromatic amino acids), PxAry1 and PxAry2 belong to the arylphorin-like protein genes. Analysis using Northern hybridization and RT-PCR showed varying levels of genes expression in the developmental stages with a small difference between sexes. Expression of both genes in fourth instar larvae was suppressed after treatment with a JH-analog. Southern hybridization revealed the presence of multiple arylphorin genes in the genome., (c 2007 Wiley-Liss, Inc.)

The rice stem borer, Chilo suppressalis, is divided into at least two ecotypes in Japan, the Shonai ecotype (SN) which is distributed in the northern part of Japan, and the Saigoku ecotype (SG) which is distributed in the southwestern region. Cold hardiness is positively correlated with the level of glycerol in both ecotypes. To investigate whether ecological distribution affects glycerol accumulation and cold hardiness development in these two ecotypes, overwintering larvae of the SN and SG ecotypes were concurrently exposed to the Shonai district. Obvious differences in the progress of glycerol accumulation and cold hardiness development in SN and SG larvae were found in early winter in the Shonai district. The levels of glycerol content and cold hardiness were low in October and high in January in both ecotypes, but those levels were different within this period (November and December) between ecotypes; the levels in SN larvae quickly reached their maximum, whereas, in SG larvae levels increased slowly. Under controlled conditions, the effect of the period of acclimation at 10 degrees C and subsequent low-temperature (5 degrees C) exposure on glycerol accumulation was investigated. These results indicated that glycerol accumulation in SN was stimulated by the progression of diapause termination, whereas a higher cumulative effect on glycerol production in SG was found when diapause was in a deep state.

This study revealed two distinct alternatively spliced exons, A1 and A2, encoding part of the domain IIS4-IIS5 of the para-sodium channel gene in the diamondback moth (DBM). Exons A1 and A2, respectively, revealed 79% and 91% identity at the nucleotide and amino acid levels. Both alternative exons included the T929I site, which has been associated with pyrethroid resistance in DBM. In the pyrethroid-resistant strain, susceptible (Thr) and resistant (Ile) amino acids were encoded at the T929I site in exons A1 and A2, respectively, but in the pyrethroid-susceptible strain, only Thr was encoded at the site in both exons. The transcripts containing exon A1 were expressed constitutively in all developmental stages. The transcripts containing exon A2 were also detected in all developmental stages, but the levels were significantly lower in the 3rd and 4th instar larvae. Tissue-specific data from the 4th instar larvae and adults showed that the expression of transcripts containing exon A2 was higher in heads than in bodies. These findings suggest that alternative splicing of the para-sodium channel gene might produce distinct channels with different sensitivities to pyrethroids, possibly in a tissue-specific manner.

The complete cDNA sequences of heat shock protein 90 (hsp90) and of heat shock cognate protein 70 (hsc70) were cloned by reverse transcription polymerase chain reaction from the rice stem borer, Chilo suppressalis Walker. They potentially encode a 717-amino-acids (hsp90) and a 652-amino-acids (hsc70) protein, with calculated molecular weight of 82.5 and 71.3 kDa, respectively. The deduced amino acid sequence of hsp90 showed the highest homology of 97.2% to Spodoptera frugiperda hsp90. The closest match of C. suppressalis hsc70 was with Manduca sexta hsc70 at 98.0% identity. Expression of hsp90 in diapausing larvae was higher than that in non-diapausing larvae. No such up-regulation in diapausing larvae was observed for hsc70. In non-diapausing larvae, but not in diapausing ones, hsp90 expression was up-regulated by cold acclimation. Hsc70 expression slightly decreased during cold acclimation irrespective of the state of diapause. Involvement of hsp90 and hsc70 in larval diapause and cold tolerance acquisition in C. suppressalis is discussed.

Heat shock protein genes, hsp90, hsc70, and hsp19.5, were cloned and sequenced from the diamondback moth, Plutella xylostella (L.) by RT-PCR and RACE method. The cDNA sequence analysis of hsp90 and hsp19.5 revealed open reading frames (ORFs) of 2,151 and 522 bp in length, which encode proteins with calculated molecular weights of 82.4 and 19.5 kDa, respectively. Analysis of cDNA from hsc70 revealed an ORF of 1,878 bp coding a protein with a calculated molecular weight of 69.3 kDa. Furthermore, the analysis of genomic DNA from hsc70 confirmed the presence of introns while no introns were apparent in hsp90 and hsp19.5. Southern blot analysis suggested the presence of multiple copies of each gene family in the DBM genome. Detectable expression of hsp19.5 was observed at the pupal stage while expression of hsp90 and hsc70 was detected at both pupal and adult stages. At adult stage, females showed a higher expression of hsp90 and hsc70 than males. An increased expression was observed in all three genes after exposure to a high temperature in both sexes. These results suggest that in addition to a heat shock response, these HSP genes might be involved in other functions during the course of development in DBM.

Overwintering larvae of the rice stem borer, Chilo suppressalis accumulate glycerol and are freezing tolerant to about -25 degrees C. However, non-diapausing larvae cannot accumulate glycerol and are killed by freezing. We compared the extent of tissue damage, the effects of glycerol concentration, and the transport of glycerol and water in fat body tissues from these larvae at selected freezing temperatures. Tissues from overwintering larvae, but not non-diapausing larvae, survive when frozen at -20 degrees C with 0.25 M glycerol, but the protection afforded by glycerol is offset by the water-channel inhibitor mercuric chloride. Glycerol in higher concentration (0.75 M) affords some protection even to the fat body of non-diapausing larvae. Radiotracer assays of overwintering larvae show that water leaves the tissues during freezing while glycerol enters, and that mercuric chloride disrupts this process. Transport is also disrupted after lethal freezing at -35 degrees C. Therefore, membrane transport of water and glycerol is involved in the avoidance of freezing injury to fat body cells of the rice stem borer, apparently by mediating the replacement of water with glycerol in freezing-tolerant tissues.

In the present study, we cloned and sequenced the entire coding regions of two glutathione S-transferase (GST) genes encoding PxGSTs and PxGSTe from the diamondback moth, Plutella xylostella L. (Lepidoptera: Yponomeutidae), along with their respective 5' and 3' flanking regions. The PxGSTs gene was composed of four exons and three introns. Based on amino acid identity with GST genes from other insects, this gene was classified as a member of the Sigma class. The gene encoding PxGSTe had an intron in the 5' flanking region. Southern blot analysis showed that PxGSTs was a single copy gene, whereas there were homologous members of the PxGSTe gene in the P. xylostella genome. RNA gel blot analysis indicated that the expression levels of both genes changed with the developmental stage of P. xylostella.

Background: Telomerase is a ribonucleoprotein enzyme that extends telomere specific repeats on the ends of chromosomes. Telomerase activity has been detected frequently in various types of human tumors and has been associated with cell immortality and oncogenesis. Human telomerase reverse transcriptase (hTERT), a telomerase catalytic subunit, reportedly regulates telomerase activity. Little is known about telomerase activity and hTERT mRNA expression in soft tissue tumors. The objective of this study was to clarify the correlation between these two parameters and clinical aggressiveness in soft tissue tumors., Methods: In 41 surgically resected soft tissue tumors, telomerase activity was measured by the fluorescence-based telomeric repeat-amplification protocol and hTERT mRNA expression was determined by real-time polymerase chain reaction., Results: Telomerase activity was detected in 52% of sarcomas and in none of the benign soft tissue tumors (P < 0.05). Telomerase activity was found in 77% of 13 locally recurrent sarcomas and in 89% of 9 sarcomas with distant metastasis. The frequency of the presence of telomerase activity in those tumors was significantly greater compared with the frequency of telomerase activity in the other sarcomas (P < 0.05 and P < 0.01, respectively). All telomerase positive sarcomas expressed hTERT mRNA. The mean level of hTERT mRNA expression in sarcomas was significantly greater compared with the mean hTERT mRNA expression level in benign tumors (P < 0.05) and in locally recurrent sarcomas compared with primary sarcomas (P < 0.001)., Conclusions: The results of the current study suggest that the detection of telomerase activity and the level of hTERT mRNA expression are useful markers for evaluating the clinical aggressiveness in soft tissue tumors., (Copyright 2002 American Cancer Society.)

We investigated the biochemical and pharmacological properties of a new adenosine A(3) receptor antagonist, KF26777 (2-(4-bromophenyl)-7,8-dihydro-4-propyl-1H-imidazo[2,1-i]purin-5(4H)-one dihydrochloride). This compound was characterized using N(6)-(4-amino-3-iodobenzyl)adenosine-5'-N-methyluronamide ([125I]AB-MECA) or [35S]guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) binding to membranes from human embryonic kidney 293 (HEK293) cells expressing human adenosine A(3) receptors. KF26777 showed a K(i) value of 0.20+/-0.038 nM for human adenosine A(3) receptors labeled with [125I]AB-MECA and possessed 9000-, 2350- and 3100-fold selectivity vs. human adenosine A(1), A(2A) and A(2B) receptors, respectively. The inhibitory mode of binding was competitive. KF26777 inhibited the binding of [35S]GTPgammaS stimulated by 1 microM 2-chloro-N(6)-(3-iodobenzyl)adenosine-5'-N-methyluronamide (Cl-IB-MECA). The IC(50) value was 270+/-85 nM; the compound had no effect on basal activity. Dexamethasone treatment for HL-60 cells, human promyelocytic leukemia, up-regulated functional adenosine A(3) receptors expression, and resulted in the enhanced elevation of intracellular Ca(2+) concentration ([Ca(2+)](i)) via the adenosine A(3) receptor. KF26777 antagonized this [Ca(2+)](i) mobilization induced by Cl-IB-MECA, with a K(B) value of 0.42+/-0.14 nM. These results indicate that KF26777 is a highly potent and selective antagonist of the human adenosine A(3) receptor.

Overwintering larvae of the Shonai ecotype of the rice stem borer, Chilo suppressalis, enter diapause in early September and terminate diapause at the end of October. Cold acclimation at 0 degrees C did not influence glycerol, trehalose or glycogen content in larvae collected on 22 September. Acclimation at 0 degrees C increased the glycerol content and reduced the glycogen content significantly in larvae collected on 2 October and 22 November compared with acclimation at 15 degrees C. These results indicate that overwintering larvae at different phases of diapause development respond differently to the low temperature stimulus for glycerol synthesis. Thus, we evaluated the metabolic rearrangements associated with glycerol synthesis during diapause development and after temperature acclimation. Larvae collected on 2 October were acclimated at 15 degrees C for 15 and 60 days. Some of those acclimated at 15 degrees C were then moved to 0 degrees C for 15 days. The larvae acclimated at 15 degrees C for 15 days were in deep diapause and accumulated little glycerol, while larvae acclimated at 15 degrees C for 60 days were nearly ready to emerge from diapause and accumulated glycerol at 155.5 &mgr;mol/g. When larvae acclimated to 15 degrees C for 15 days were transferred to 0 degrees C, glycerol accumulation was stimulated to the same extent (ca 140 &mgr;mol/g) as it was in larvae that were acclimated to 15 degrees C for 60 days and then transferred to 0 degrees C. These results indicate that low temperature has a cumulative effect on glycerol production in larvae at different phases of diapause development. Glycerol accumulation was accomplished by activation of glycogen phosphorylase and inhibition of fructose-1,6-bisphosphatase, and activation of enzymes associated with glycerol synthesis, mainly glyceraldehyde-3-phosphatase and polyol dehydrogenase with glyceraldehyde activity.

The heteropteran Pyrrhocoris apterus (L.) does not survive freezing of its body fluids; there is a good correlation between values of survival at subzero temperatures and the supercooling point (SCP), i.e., the temperature at which body fluids start to freeze. The decrease of the SCP and thus the increase in cold hardiness is regulated by photoperiod and temperature. The relative importance of these factors depends on the physiological state of the insect. The SCP is about -7&deg;C at the onset of prediapause and a decrease of about 4-5&deg;C is associated with the development of the diapause syndrome in adults; these processes both are induced by a short-day photoperiod with temperature playing a secondary role. The induction of the diapause syndrome is a prerequisite for the subsequent decrease of the SCP by about 5-6&deg;C during cold acclimation. An intermediate temperature of 15&deg;C, or fluctuating outdoor temperatures and short-day photoperiods, are more suitable for the decrease of SCP than 5&deg;C in continuous darkness. The sensitivity to photoperiod gradually disappears during the development of diapause; after the termination of diapause around the winter solstice the SCP irreversibly increases at a high temperature of 26&deg;C even if exposed to a short-day photoperiod. The SCPs of hemolymph, gut, fat body, and gonads were compared to whole-body SCP. The gut was identified as the primary site of ice nucleation because its SCP value was very similar to the value for the whole body in both short-day and long-day insects. The SCPs of other organs, including the hemolymph, were always lower than the whole body SCP. Food was not a source of ice nucleating agents because the SCP of freshly ecdysed adults remained high after 2 weeks of starvation. In contrast, feeding was a prerequisite for the decrease of the SCP during prediapause. In postdiapause insects, the SCP increased at high temperatures in spite of the absence of food.

The effects of culture conditions on the ice nucleus production of Fusarium moniliforme var. subglutinans isolated from the gut of larvae of the rice stem borer (Chilo suppressalis Walker) were examined. The ice nucleus production was only affected by cultivation temperature and pH: the optimum temperature and pH were 15 degrees C to 20 degrees C and 4.0 to 6.0, respectively.